Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers.

A method for apolipoprotein (apo) E genotyping was developed using the polymerase chain reaction (PCR) with allele-specific oligonucleotide primers (ASP). Synthetic oligonucleotides with base-pair mismatches at the 3' terminus were used as primers to amplify the apoE gene in subjects previously...

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Main Authors: BF Main, PJ Jones, RT MacGillivray, DK Banfield
Format: Article
Language:English
Published: Elsevier 1991-01-01
Series:Journal of Lipid Research
Online Access:http://www.sciencedirect.com/science/article/pii/S0022227520422576
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author BF Main
PJ Jones
RT MacGillivray
DK Banfield
author_facet BF Main
PJ Jones
RT MacGillivray
DK Banfield
author_sort BF Main
collection DOAJ
description A method for apolipoprotein (apo) E genotyping was developed using the polymerase chain reaction (PCR) with allele-specific oligonucleotide primers (ASP). Synthetic oligonucleotides with base-pair mismatches at the 3' terminus were used as primers to amplify the apoE gene in subjects previously phenotyped using isoelectric focusing (IEF). Complementary primer-allele combinations were specifically amplified by PCR, together with a control pair of primers specific to the human prothrombin gene. Identification of genotype by PCR using ASP was consistent with the phenotypes that were determined by IEF for 14 healthy normolipidemic subjects. These results were achieved using DNA isolated from buccal epithelial cells obtained from a mouthwash or DNA extracted from leukocytes. Genotype identification required analysis of the PCR products on an ethidium-stained agarose gel, yielding results 3 h after DNA extraction. In comparison with other current methods, PCR using ASP is suggested as a rapid and simple noninvasive technique for determining population apoE allelic distribution.
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spelling doaj.art-0221ed42ac5e44858477b3fd0bad54a32022-12-21T22:35:53ZengElsevierJournal of Lipid Research0022-22751991-01-01321183187Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers.BF Main0PJ Jones1RT MacGillivray2DK Banfield3Division of Human Nutrition, University of British Columbia, Vancouver, Canada.Division of Human Nutrition, University of British Columbia, Vancouver, Canada.Division of Human Nutrition, University of British Columbia, Vancouver, Canada.Division of Human Nutrition, University of British Columbia, Vancouver, Canada.A method for apolipoprotein (apo) E genotyping was developed using the polymerase chain reaction (PCR) with allele-specific oligonucleotide primers (ASP). Synthetic oligonucleotides with base-pair mismatches at the 3' terminus were used as primers to amplify the apoE gene in subjects previously phenotyped using isoelectric focusing (IEF). Complementary primer-allele combinations were specifically amplified by PCR, together with a control pair of primers specific to the human prothrombin gene. Identification of genotype by PCR using ASP was consistent with the phenotypes that were determined by IEF for 14 healthy normolipidemic subjects. These results were achieved using DNA isolated from buccal epithelial cells obtained from a mouthwash or DNA extracted from leukocytes. Genotype identification required analysis of the PCR products on an ethidium-stained agarose gel, yielding results 3 h after DNA extraction. In comparison with other current methods, PCR using ASP is suggested as a rapid and simple noninvasive technique for determining population apoE allelic distribution.http://www.sciencedirect.com/science/article/pii/S0022227520422576
spellingShingle BF Main
PJ Jones
RT MacGillivray
DK Banfield
Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers.
Journal of Lipid Research
title Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers.
title_full Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers.
title_fullStr Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers.
title_full_unstemmed Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers.
title_short Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers.
title_sort apolipoprotein e genotyping using the polymerase chain reaction and allele specific oligonucleotide primers
url http://www.sciencedirect.com/science/article/pii/S0022227520422576
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